각막내피손상 및 비손상 모델에서 각막내피세포와 각막기질의 상호작용
Interactions of Corneal Endothelial Cells with Stromal Cells during Corneal Endothelial Injury
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목적: 화학적으로 유발된 각막내피손상의 치유와 세포 배양을 이용하여 각막내피세포와 기질세포의 상호작용을 조사하였다. 대상과 방법: 25마리의 토끼를 NaOH에 각막내피가 손상된 군과 대조군 두 그룹으로 나누었다. 0.05 N NaOH 0.1 mL를 전방에 10분동안 처리하여 각막내피손상을 유발한 후 1일, 1주, 2주 및 4주에 임상적으로 평가하였다. 각막내피세포와 각막기질세포를 배양하여Transwell system에서 공배양과 그렇지 않을 때의 각막내피의 상처 치유 속도와 배양액 내의 사이토카인을 분석하였다. 결과: 각막부종과 불투명도 점수는 대조군보다 NaOH에 손상된 모든 토끼에서 더 높았다. 1일째에 NaOH에 각막내피세포가 손상된토끼에서 각막기질세포에는 영향을 거의 주지 않으면서 각막내피세포에 apoptosis가 관찰되었다. 손상 2주 후, 각막내피세포가 주변부에서 중심부로 이동하였다. 각막기질세포는 각막내피세포의 존재와 함께 데스메막에 평행하게 정렬되었다. 배양각막내피세포의 상처 치유는 각막기질세포와의 공배양에서 빨랐고 배양액 내로의 transforming factor-β1 및 interleukin-6 분비는 각막내피세포 또는각막기질세포의 개별배양에서보다 공배양에서 더 낮았다. 결론: 전방 내 NaOH 주입은 기질 조직에 최소한의 영향을 미치는 각막내피손상의 모델이 될 수 있을 것으로 보인다. 또한, 상처 치유과정에서 각막내피세포와 각막기질세포 사이의 상호작용은 각막내피손상의 치유 과정에 필요한 사이토카인 생산과 관련이 있다.
Abstract
Purpose: To investigate the interaction of corneal endothelial cells (CECs) with corneal stromal cells (CSCs) on the healing of chemically induced corneal endothelial injury. Methods: Twenty-five rabbits were divided into two groups: rabbits exposed to NaOH and controls. Rabbits were clinically evaluated and sacrificed at 1 day, 1 week, 2 weeks, and 4 weeks after endothelial injury. Corneal endothelial cells and corneal stromal cells were cultured and the wound healing rate of the corneal endothelium and cytokines in the culture medium were analyzed with and without co-culture in the Transwell system. Results: Corneal edema and opacity scores were higher in all rabbits exposed to NaOH than in controls. Corneal endothelial apoptosis with little effect on CSCs was observed in rabbits exposed to NaOH at day 1. Two weeks after injury, CECs migrated from the periphery. CSCs were aligned in parallel to Descemet's membrane, which exhibited the presence of CECs. Transforming factor- β1 and interleukin-6 secretion was lower in the co-culture system than in individual cultures of CECs or CSCs. Conclusions: These results revealed intracameral NaOH injection is a model of corneal endothelial injury with minimal impact on the stroma. Communication between CECs and CSCs during wound healing may lead to the modulation of cytokine production, which is necessary for an enhanced healing process of corneal endothelial injury.
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